- PA5-21368 - Invitrogen Antibodies PAX8 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of PAX8C using 30 µg of 293T lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a PAX8C polyclonal antibody (Product # PA5-21368) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of PAX8 was performed by separating 30 µg of 293T whole cell and nuclear extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a PAX8 Polyclonal Antibody (Product # PA5-21368) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of PAX8 was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a PAX8 Polyclonal Antibody (Product # PA5-21368) at a dilution of 1:5000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-PAX8 Polyclonal Antibody (Product # PA5-21368) and a 57 kDa band corresponding to PAX8 was observed across the cell lines tested except IMR-32 and Jurkat. An uncharacterized band of ~17 kDa was also observed in positive cell lines. Modified whole cell extracts (1% SDS) (30 µg lysate) of SK-OV-3 (Lane 1), OVCAR-3 (Lane 2), MCF7 (Lane 3), PC-3 (Lane 4), HEK-293 (Lane 5), HeLa (Lane 6), IMR-32 (Lane 7) and Jurkat (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036), using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of PAX8C in paraformaldehyde-fixed A549 cells using a PAX8C polyclonal antibody (Product # PA5-21368) at a 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of paraformaldehyde-fixed A549, using PAX8 (Product # PA5-21368) antibody at 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of PAX8 was performed using 70% confluent log phase SK-OV-3 cells. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. The cells were labeled with PAX8 Polyclonal Antibody (Product # PA5-21368) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27034, 1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents IMR-32 cells having no expression of PAX8. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Paraffin) analysis of PAX8 was performed in paraffin-embedded DLD1 xenograft tissue using PAX8 Polyclonal Antibody (Product # PA5-21368) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Paraffin) analysis of PAX8 was performed in paraffin-embedded mouse liver tissue using PAX8 Polyclonal Antibody (Product # PA5-21368) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: PAX8 Polyclonal Antibody detects PAX8 protein at cytosol and nucleus on rat thyroid gland by immunohistochemical analysis. Sample: Paraffin-embedded rat thyroid gland. PAX8 Polyclonal Antibody (Product # PA5-21368) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: PAX8 antibody immunoprecipitates PAX8 protein-DNA in ChIP experiments. ChIP Sample: 293T whole cell lysate/extract. A: 5 µg preimmune rabbit IgG. B: 5 µg of PAX8 antibody (Product # PA5-21368). The precipitated DNA was detected by PCR with primer set targeting to E2F1 promoter.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: PAX8 Polyclonal Antibody immunoprecipitates PAX8 protein in IP experiments. IP Sample: 1,000 µg 293T whole cell lysate/extract A. 40 µg 293T whole cell lysate/extract B. Control with 2 µg of preimmune rabbit IgG C. Immunoprecipitation of PAX8 protein by 2 µg of PAX8 Polyclonal Antibody (Product # PA5-21368) 10% SDS-PAGE The immunoprecipitated PAX8 protein was detected by PAX8 Polyclonal Antibody (Product # PA5-21368) diluted at 1:1,000.

PA5-21368